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Information × Registration Number 0217U000631, 0113U003764 , R & D reports Title The investigation of ultrafast cooling of biological objects to create a device for cryopreservation popup.stage_title Head Sydorenko Myhail V., Registration Date 16-02-2017 Organization Department of Biotechnical Problems of Diagnostics, Institute of Problems of Cryobiology and Cryomedicine of NAS of Ukraine popup.description2 Abstract. Research report: 64 p. 20 Fig. 15 of Table. 67, literature sources. The object of study: MCF-7 cell adenocarcinoma of the breast, colon cancer cells HT29, cryoprotective agents. Objective: To investigate the biological and chemical characteristics of the phenomenon of ultra-rapid cooling of biological objects to create and test the device layout, which provides a reduction in temperature of the object at a speed of up to 300 ° C / min. The development of cryopreservation protocols simple biological objects using the vitrification method. The main objectives of the work. Creation of active-device layout for quick cooling of biological objects of varying degrees of complexity. Preparation of technical documentation for the device for the ultra-fast freezing. Determination of temperature gradients on the verge of freezing transitions "nitrogen" - "the inner surface of the container" - "biological object", for different compositions of cryoprotectants. Selection of the composition of cryoprotectants to increase the efficiency of cryopreservation of biological objects of various sizes and degrees of complexity using the device layout. Research methods and instruments. To create a mock-unit developments were used Experimental Factory of the Institute of Cryobiology and Cryomedicine problems of NAS of Ukraine. To measure the temperature of freezing regime used thermocouples and method developed by members of the Experimental Factory IPKiK NAS of Ukraine software. Vitrifikatsinnye solutions (BP) developed by staff of the Laboratory of marker compounds VBTPD IPCC NAS of Ukraine consisted of the culture medium, serum, of DMSO, sucrose, glycerol, ethylene glycol, polyethylene glycol, CMC in different proportions. When adding BP used five different protocols at a slow and super-quick freezing apparatus using mock sample. To determine the effectiveness of the freezing protocols used standard cytological methods: MTT test, dye trypan blue, light microscopy. The results and their novelty. As a result of the work it was created and launched the current layout of the device for ultrafast cooling of biological objects. With this device kinetics Vitrifikatsiynih temperature changes in solutions of different composition and temperature gradients on the edge transitions freezing "nitrogen" - "internal surface of the container" - "bioobject" revealed a correlation between the molecular weight (MW) of the variable speed component and cryoprotectants. lowering the temperature in the cryo-container sign of the correlation coefficient between MM cryoprotectant and temperature reduction rate in the range from 0 to 40 pre-positive (r = 0, 8144), while in the range from -40 to -. 170 ° C - negative (r = -. 0.9096) the viability of data cells using MTT assay also indicate a strong negative correlation cell survival after freezing/thawing cycles and the amount of cryoprotectant molecules in the range of 342-20000 Dalton, r = - 0,9908 from the data as well. it follows that the presence of high-molecular substances cryoprotective mixtures leads to smoothing the curve of temperature versus time and to decrease the cooling velocity in comparison with the sucrose solution. At the same time, low-molecular PEG (1,5 kDa) cryoprotectant was more effective than the high molecular weight PEG, and sucrose. Addition of PEG to low molecular weight mixtures cryoprotective enhances cell survival by 1.3 times without sucrose and 2.12 times - with sucrose. Furthermore, the presence of sucrose provides more efficient freezing conditions. Indicator cell survival with most BP increased by adding carboxymethyl cellulose (CMC). According to the research, two registered patent: "Method for superfast freezing of human somatic cells" and "composition of the medium to ultra-fast freezing of human somatic cells”. Key words: cryopreservation, a device for ultra-rapid cooling, polyethylene glycol, CMC. Product Description popup.authors Безуглий С.В. Бородай Н.В. Бурлака Дмитро Петрович Сидоренко Михайло Васильвич Угнівенко А.П. Якимчук О.М. popup.nrat_date 2020-04-02 Close